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Published on: June 6, 2020
[Screening of interacting proteins of idiopathic gonadotropin-releasing hormone deficiency pathogenic gene RNF216]
Wenting Dai1, Zuiming Jiang, Min Gu
1Center of Clinical Laboratory, the Affiliated Zhuzhou Hospital of Xiangya Medical College, Central South University, Zhuzhou, Hunan 412007, China. 152501019@csu.edu.cn.
Objective:
To screen proteins interacting with ring finger protein 216(RNF216) through yeast two hybrid experiment, and further clarify the role of RNF216 in the pathogenesis of gonadotropin-releasing hormone deficiency.
Methods:
A recombinant expression vector pGBKT7-RNF216 was constructed and transformed into yeast Y2HGold, which was hybridized with a human cDNA library in order to screen proteins interacting with RNF216. The interaction was verified in yeast Y2HGold.
Results:
A recombinant expression vector pGBKT7-RNF216 was successfully constructed and expressed in yeast Y2HGold. Filamin B (FLNB) was identified by yeast two hybrid experiment, and their interaction was verified in yeast Y2HGold.
Conclusion:
An interaction between FLNB and RNF216 was identified through yeast two hybrid experiment. RNF216 may affect the proliferation and migration of GnRH neurons by regulating FLNB or FLNB/FLNA heterodimers.
Insights
Researchers identified Filamin B (FLNB) interacting with ring finger protein 216 (RNF216) using yeast two-hybrid screening. This finding may clarify RNF216
Area of Science:
- Molecular Biology
- Genetics
- Neuroendocrinology
Background:
- Ring finger protein 216 (RNF216) is implicated in various cellular processes.
- The precise role of RNF216 in the pathogenesis of gonadotropin-releasing hormone (GnRH) deficiency remains unclear.
- Understanding protein interactions of RNF216 is crucial for elucidating its function in neuroendocrine disorders.
Purpose of the Study:
- To identify proteins that interact with RNF216 using a yeast two-hybrid system.
- To investigate the potential role of RNF216 in the development of GnRH deficiency.
Main Methods:
- Construction and yeast transformation of the pGBKT7-RNF216 expression vector.
- Yeast two-hybrid screening against a human cDNA library to identify RNF216-interacting proteins.
- Verification of identified interactions in the yeast Y2HGold system.
Main Results:
- Successful construction and expression of the pGBKT7-RNF216 vector in yeast.
- Identification of Filamin B (FLNB) as a protein interacting with RNF216.
- Confirmation of the RNF216-FLNB interaction through yeast two-hybrid experiments.
Conclusions:
- A direct interaction between FLNB and RNF216 was established.
- RNF216 may influence GnRH neuron proliferation and migration.
- This regulation could occur via FLNB or FLNB/FLNA heterodimers, offering insights into GnRH deficiency pathogenesis.
